Immunofluorescence analysis of Proteasome 20S Subunit alpha-5 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PSMA5 Rabbit Polyclonal (PA11962) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Hamster, Human, Mouse, Rat|
|Published species reactivity||Rat, Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C A(120) L Q F G E E D A D P G A M(133) of human proteasome 20S subunit alpha 5.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||1 - 3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-1962 detects proteasome 20S subunit alpha 5 from human, rat, mouse and hamster cell lysates.
PA1-1962 has been successfully used in Western blot and immunofluorescence procedures. By Western blot, this antibody detects a 26.5 kDa protein representing proteasome 20S subunit alpha 5 from human, rat, mouse, and hamster cell extracts.
PA1-1962 immunogen is a synthetic peptide corresponding to residues C A(120) L Q F G E E D A D P G A M(133) of human proteasome 20S subunit alpha 5. PA1-1962 immunizing peptide (Cat. # PEP-210) is available for use in neutralization and control experiments.
Proteolytic degradation is critical to the maintenance of appropriate levels of short-lived and regulatory proteins as important and diverse as those involved in cellular metabolism, heat shock and stress response, antigen presentation, modulation of cell surface receptors and ion channels, cell cycle regulation, transcription, and signaling factors. The ubiquitin-proteasome pathway deconstructs most proteins in the eukaryotic cell cytosol and nucleus. Others are degraded via the vacuolar pathway which includes endosomes, lysosomes, and the endoplasmic reticulum.
The 26S proteasome is an ATP-dependent, multisubunit (~31), barrel-shaped molecular machine with an apparent molecular weight of ~2.5 MDa. It consists of a 20S proteolytic core complex which is crowned at one or both ends by 19S regulatory subunit complexes. The 19S regulatory subunits recognize ubiquitinated proteins and play an essential role in unfolding and translocating targets into the lumen of the 20S subunit. An enzymatic cascade is responsible for the attachment of multiple ubiquitin molecules to lysine residues of proteins targeted for degradation. Several genetic diseases are associated with defects in the ubiquitin-proteasome pathway. Some examples of affected proteins include those linked to cystic fibrosis, Angelman and quote;s syndrome, and Liddle syndrome.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The transition zone protein Rpgrip1l regulates proteasomal activity at the primary cilium.
PA1-1962 was used in immunocytochemistry to use knockout mice to determine the function of RPGRIP1L.
|Gerhardt C,Lier JM,Burmühl S,Struchtrup A,Deutschmann K,Vetter M,Leu T,Reeg S,Grune T,Rüther U||The Journal of cell biology (210:115)||2015|
Urocortin trafficking in cerebral microvessel endothelial cells.
PA1-1962 was used in immunocytochemistry to investigate the mechanism of urocortin endocytosis in cerebral microvessel endothelial cells
|Tu H,Kastin AJ,Bjorbaek C,Pan W||Journal of molecular neuroscience : MN (31:171)||2007|
ATM regulates proteasome-dependent subnuclear localization of TRF1, which is important for telomere maintenance.
PA1-1962 was used in western blot to study the role of ATM in telomere function
|McKerlie M,Lin S,Zhu XD||Nucleic acids research (40:3975)||2012|
Neuronal induction of the immunoproteasome in Huntington's disease.
PA1-1962 was used in western blot to investigate the role of the ubiquitin-proteasome system during the neurodegeneration of huntington disease.
|Díaz-Hernández M,Hernández F,Martín-Aparicio E,Gómez-Ramos P,Morán MA,Castaño JG,Ferrer I,Avila J,Lucas JJ||The Journal of neuroscience : the official journal of the Society for Neuroscience (23:11653)||2003|